Fingerprint-based polyphenol compound determination method and system

Abstract

The invention discloses a polyphenol compound determination method based on a fingerprint spectrum, which comprises the following steps: determining chromatographic conditions of polyphenol compounds in a detection sample, and determining the content of the polyphenol compounds, the method comprises the following steps: establishing fingerprints of the same sample from a plurality of different regions by using an HPLC-ECD (High Performance Liquid Chromatography-Error Code Division) method, carrying out regional calibration on polyphenol compounds to obtain regional calibration values, carrying out first similarity and clustering analysis, and establishing a determination model for distinguishing the same sample from different producing areas; establishing HPLC-ECD fingerprints of different types of samples, carrying out variety calibration on polyphenol compounds to obtain a variety calibration value, carrying out second similarity and clustering analysis, and establishing a determination model for distinguishing different types of samples; specifying a sample type, establishing an adulterated sample fingerprint spectrum after adding different mass fractions of fake products, determining polyphenol compounds, performing third similarity and clustering analysis, and establishing an adulterated sample identification model. The invention further discloses a corresponding system, electronic equipment and a computer readable storage medium.

Description

Technical Field

[0001] The present invention relates to the technical field of fingerprint and compound determination, and in particular to a method and system for determining polyphenol compounds based on fingerprint. Background Art

[0002] Most of the antioxidant components in honey are composed of polyphenolic compounds. The content of polyphenolic compounds in honey is generally 6000ug / kg. At present, a variety of polyphenolic compounds such as caffeic acid, chlorogenic acid, protocatechuic acid, and calendulin have been detected in honey. Honey has different plant sources, and the polyphenolic compounds it contains are also very different. Therefore, polyphenolic substances not only have the function of neutralizing free radicals, but can also be used as a suitable indicator for identifying the plant source and quality control of honey. Polyphenolic compounds are generally divided into flavonoids, phenolic acids and tannins, among which flavonoids and phenolic acids are more widely present. Polyphenolic compounds are one of the main components that make honey have antioxidant activity. Honey from different plant sources contains different types of polyphenolic compounds, so polyphenolic compounds can also be used as an important indicator for evaluating honey quality and identifying honey authenticity.

[0003] The HPLC analysis method has many advantages, such as good selectivity, fast analysis speed, high separation efficiency, high sensitivity, low sample consumption, and wide application range. However, it has limitations, such as having to rely on known standards for comparison and poor analysis of isomers. The detector has certain restrictions on the sample to be tested and the mobile phase. There are certain limitations in practical application. In addition, the existing determination method has a good identification effect on honey with added sugar substances, but cannot identify other adulteration situations. Summary of the invention

[0004] The purpose of the present invention is to solve the problems existing in the prior art and propose a method and system for determining polyphenolic compounds based on fingerprints. The method of high performance liquid chromatography combined with electrochemical detection has high sensitivity and can also achieve good quantification for trace amounts of polyphenolic compounds in honey. The polyphenolic compounds in honey are the main components that make it have antioxidant activity. By detecting the polyphenolic compounds in honey, the quality of honey can be effectively evaluated, providing a scientific and quantitative basis for honey quality control. In addition, the constructed honey fingerprint can well reflect the content and difference of antioxidant components in different honeys and evaluate the quality of honey. The honey fingerprint constructed by combining similarity calculation and cluster analysis has not been recorded in the prior art. The determination of polyphenolic compounds by this method has opened up a new way for honey adulteration identification and quality control.

[0005] The first aspect of the present invention is to provide a method for determining polyphenolic compounds based on fingerprint, comprising:

[0006] S1, determining the chromatographic conditions for detecting polyphenolic compounds in a sample, and determining the content of the polyphenolic compounds;

[0007] S2, based on the determined content of polyphenolic compounds, using HPLC-ECD method to establish fingerprints of the same sample from multiple different regions, by analyzing the differences in the fingerprints of the same sample, regional calibration of polyphenolic compounds is performed on the sample to obtain regional calibration values, the regional calibration values ​​are subjected to first similarity and cluster analysis, and a determination model for distinguishing the same samples from different origins is established;

[0008] S3, establishing HPLC-ECD fingerprints of different types of samples, performing type calibration of polyphenol compounds on the samples by analyzing the differences in the HPLC-ECD fingerprints of the different types of samples to obtain type calibration values, and performing a second similarity and cluster analysis on the type calibration values, thereby establishing a determination model for distinguishing different types of samples;

[0009] S4, specify the sample type, establish the fingerprint of the adulterated sample after adding different mass fractions of counterfeit products into it, analyze the differences in the fingerprints of the adulterated samples, measure the polyphenol compounds, and then perform the third similarity and cluster analysis on the measured values ​​to establish the adulterated sample identification model.

[0010] Preferably, the S1 comprises:

[0011] S11, prepare experimental materials and experimental instruments;

[0012] S12, preparing standard substances and samples to be tested;

[0013] S13, select and optimize chromatographic conditions;

[0014] S14, conducting a determination experiment of the content of polyphenol compounds, including a preliminary determination experiment and a main determination experiment.

[0015] Preferably, the preliminary assay experiment includes a precision experiment, a repeatability experiment and a stability experiment; the main assay experiment is used to calculate a standard curve, and to determine the polyphenol compounds in the sample to be tested based on the standard curve.

[0016] Preferably, the performing of a first similarity and cluster analysis on the regional calibration values ​​to establish a determination model for distinguishing the same samples from different origins comprises:

[0017] The peak time of the element with the highest content in each sample is in the middle of the entire chromatogram, the peak shape is stable and there is no influence of other peaks, and the peak corresponding to the element is selected as the reference peak, that is, the internal standard peak;

[0018] Calculating the normalized areas of other multiple common peaks based on the reference peak;

[0019] Classifying the samples according to the normalized areas and forming a matrix;

[0020] The similarity between relative peak areas was calculated and classified, and a determination model for distinguishing the same samples from different origins was established based on the classification.

[0021] Preferably, the performing a second similarity and cluster analysis on the type calibration values ​​to establish a measurement model for distinguishing different types of samples comprises:

[0022] Analyzing the variation trend of the chromatographic peak areas of samples from different sources and regions to obtain a first analysis result, wherein the first analysis result is used to characterize the variation of the content of polyphenolic compounds in different types of samples;

[0023] Analyzing the sample similarity to obtain a second analysis result, wherein the second analysis result is used to identify the sample type by comparing the sample similarity according to a predetermined condition threshold and a standard spectrum;

[0024] Performing cluster analysis on the obtained type calibration values ​​to obtain a third analysis result, wherein the third analysis result is used to distinguish the origin and type of a large number of samples;

[0025] A measurement model for distinguishing different types of samples is established based on the first analysis result, the second analysis result, and the third analysis result.

[0026] Preferably, the method of performing a third similarity and cluster analysis on the measured values ​​to establish an adulterated sample identification model comprises:

[0027] Establish standard fingerprints;

[0028] Collect data from 3 to 32 minutes when the chromatogram is eluted, and calculate the similarity between the sample to be tested and the standard fingerprint. The closer the similarity, the more similar the two samples are, and similar analysis results are obtained;

[0029] The peak time of the element with the highest content in each sample is in the middle of the entire chromatogram, the peak shape is stable and there is no influence of other peaks, and the peak corresponding to the element is selected as the reference peak, that is, the internal standard peak;

[0030] Perform interpolation matching on all common peaks other than the reference peak to be clustered, that is, fill in the missing data with a value of 0, and mark the missing time point with the retention time of the peak position;

[0031] The normalized areas of the common peaks other than the reference peak were calculated, and the obtained results were clustered using SPSS to obtain the cluster analysis results;

[0032] A model for identifying adulterated samples is established based on the similarity analysis results and the cluster analysis results.

[0033] Preferably, the establishment of the standard fingerprint spectrum includes:

[0034] The chromatographic information of the sample to be tested is obtained by the HPLC-ECD method;

[0035] The component corresponding to each chromatographic peak is determined according to the retention time recorded in the chromatographic information, and the content of the component can be calculated according to the peak area;

[0036] The commonality of samples of the same type, i.e., common peaks, is found through retention time, and the similarity between samples is evaluated through the relative peak areas between the common peaks;

[0037] The map information containing the common characteristics of the species is used as the standard fingerprint map.

[0038] The second aspect of the present invention is to provide a polyphenol compound determination system based on fingerprint spectrum, comprising:

[0039] A polyphenol compound determination module (101) is used to determine the chromatographic conditions for detecting polyphenol compounds in a sample and to determine the content of polyphenol compounds;

[0040] The first model building module (102) is used to establish fingerprints of the same sample from multiple different regions by using the HPLC-ECD method based on the determined content of polyphenolic compounds, perform regional calibration of polyphenolic compounds on the same sample by analyzing the differences in the fingerprints of the same sample to obtain regional calibration values, perform first similarity and cluster analysis on the regional calibration values, and establish a determination model for distinguishing the same samples from different origins;

[0041] The second model building module (103) is used to establish HPLC-ECD fingerprints of samples of different types, analyze the differences in the HPLC-ECD fingerprints of samples of different types, perform type calibration on the polyphenol compounds, and obtain type calibration values, and perform a second similarity and cluster analysis on the type calibration values, thereby establishing a determination model for distinguishing samples of different types;

[0042] The third model building module (104) is used to specify the sample type, establish the fingerprint of the adulterated sample after adding different mass fractions of counterfeit products thereto, analyze the differences in the fingerprint of the adulterated sample, measure the polyphenol compounds thereon, and perform the third similarity and cluster analysis on the measured values, thereby establishing an adulterated sample identification model.

[0043] A third aspect of the present invention provides an electronic device, comprising a processor and a memory, wherein the memory stores a plurality of instructions, and the processor is configured to read the instructions and execute the method described in the first aspect.

[0044] A fourth aspect of the present invention provides a computer-readable storage medium, wherein the computer-readable storage medium stores a plurality of instructions, and the plurality of instructions can be read by a processor and execute the method described in the first aspect.

[0045] Beneficial effects of the method and system of the present invention:

[0046] (1) The chromatographic conditions for detecting polyphenols in honey were optimized, and the contents of ten polyphenols in sunflower honey and rapeseed honey were determined.

[0047] (2) The HPLC-ECD method was used to establish the fingerprint of the same type of honey from multiple different regions. The polyphenol compounds were determined by analyzing the differences in the fingerprints of the same type of honey. The measured values ​​were then subjected to first similarity and cluster analysis to establish an effective method for distinguishing the same type of honey from different origins.

[0048] (3) Establish HPLC-ECD fingerprints of different types of honey, determine the polyphenolic compounds by analyzing the differences in the HPLC-ECD fingerprints of different types of honey, and then perform a second similarity and cluster analysis on the measured values, thereby establishing an effective method for distinguishing different types of honey.

[0049] (4) A type of honey is designated, and a fingerprint of the adulterated honey is established after adding different mass fractions of rice syrup into the honey. The differences in the fingerprints of the adulterated honey are analyzed, and the polyphenol compounds are determined. The measured values ​​are subjected to third similarity and cluster analysis, thereby establishing a method for identifying adulterated honey. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the related technologies, the drawings required for use in the specific embodiments or the related technical descriptions will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0051] Figure 1 A flow chart of a method for determining polyphenolic compounds based on fingerprints according to an embodiment of the present invention;

[0052] Figure 2 It is a flow chart of step S1 in the method for determining polyphenol compounds based on fingerprint spectrum provided in an embodiment of the present invention;

[0053] Figure 3 A schematic diagram of the architecture of a polyphenol compound determination system based on fingerprint spectrum provided according to an embodiment of the present invention;

[0054] Figure 4A structural diagram of an electronic device provided according to an embodiment of the present invention. DETAILED DESCRIPTION

[0055] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0056] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0057] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0058] Embodiment 1 (using honey as a sample example for actual implementation)

[0059] like Figure 1 As shown, this embodiment provides a method for determining polyphenol compounds based on fingerprint, comprising:

[0060] The purpose of establishing a fingerprint is to determine the similarity of chemical components in the chromatograms of different samples to be tested, and to distinguish the samples to be tested by comparing them with the standard. This requires quantifying the chromatogram information into digital information that can accurately define and measure the samples to be tested, and a mathematical method that can calculate the similarity of the chromatograms. In the analysis of chromatograms, peak purity and peak matching are two key points. Because chromatographic instruments are complex, slightly different conditions will produce very different results. In the process of peaking the samples to be tested, it is also possible that the desired components and results will not be obtained. This requires analysis of the peak purity and peak matching of the chromatogram, which is the key to ensuring the accuracy of the information in the construction of the fingerprint.

[0061] S1, determining the chromatographic conditions for detecting polyphenolic compounds in honey, and determining the content of polyphenolic compounds;

[0062] See also Figure 2 As a preferred implementation, S1 includes:

[0063] S11, prepare experimental materials and experimental instruments;

[0064] In this embodiment:

[0065] The experimental materials were: sunflower honey, rape honey, gallic acid, protocatechuic acid, caffeic acid, coumaric acid, ferulic acid, rutin, myricetin, calendulin, kaempferol and galangin, all purchased from Sigma; methanol (chromatographic grade); glacial acetic acid (C2H4O2) were all analytical grade;

[0066] The experimental instruments were: Agilent 1100 high performance liquid chromatograph (Agilent, USA), including a quaternary gradient pump (G1311A), a diode array detector (G1315A), an electrochemical detector (HP1049A), and an HP chromatography data acquisition workstation; HH-2 digital display constant temperature water bath (Guohua Electric Co., Ltd.); D-9143B-1 electric constant temperature blast drying oven (Shanghai Foma Experimental Equipment Co., Ltd.); electronic analytical balance (Beijing Sartorius Balance Co., Ltd.); KQ-100B ultrasonic cleaner; Milli-QG ultrapure water instrument (Millipore, USA).

[0067] S12, preparing standard substances and samples to be tested;

[0068] In this embodiment:

[0069] Preparation of standard products includes: accurately weighing 0.32 mg gallic acid, 0.3 mg protocatechuic acid, 0.35 mg caffeic acid, 0.3 mg p-coumaric acid, 0.37 mg ferulic acid, 0.42 mg rutin, 0.06 mg myricetin, 0.35 mg calendulin, 0.35 mg kaempferol, and 0.33 mg galangin in a volumetric flask, dissolving them with methanol (chromatographic grade) and dissolving them to 10 ml, shaking them to fully dissolve; taking a certain amount of each of the 10 standard products, mixing them, diluting them into 6 different concentration gradients, and storing the above-configured standard products and standard products of different concentrations in a 4°C refrigerator for later use.

[0070] The preparation of the samples to be tested includes: weighing 2 g of each of 6 types of honey from different origins or different packaging, dissolving them in ultrapure water, making up to volume with a 10 ml volumetric flask, shaking and marking the corresponding sample numbers, nectar plants, origins and collection times, storing them in a 4°C refrigerator, and taking an appropriate amount and filtering them with a 0.45 um micromembrane before sampling.

[0071] S13, select and optimize chromatographic conditions;

[0072] In this embodiment:

[0073] Chromatographic conditions are as follows: American Zorbax SB-C18 chromatographic column; mobile phase is methanol (B)-2% acetic acid water (A); gradient elution: 0-6min2%-8% B, 6-10min8%-15% B, 10-15min15%-35% B, 15-20min35%-55% B, 20-25min55%-65% B, 25-30min65%-70% B, 30-35min70%-75% B, flow rate is 1.0 ml / min;

[0074] The potential of the electrochemical detector (ECD) was 0.8 V; the detection wavelength of the diode array detector (DAD) was 360 nm; the column temperature was 30 °C; the retention times of gallic acid, protocatechuic acid, caffeic acid, p-coumaric acid, ferulic acid, rutin, myricetin, quercetin, kaempferol and galangin were 3.169 min, 5.927 min, 13.714 min, 17.182 min, 8.193 min, 19.754 min, 20.755 min, 22.717 min, 24.466 min and 28.306 min, respectively.

[0075] Reasons for optimization and optimized chromatographic conditions: Polyphenols have a good response on the electrochemical detector, and the ten phenolic acid components can be well separated, with a large response peak area and a stable peak shape. In order to obtain an effective, accurate and optimal chromatographic fingerprint, the chromatograms under various chromatographic parameters were tested and compared with reference to the experimental conditions for the determination of four polyphenols in citrus honey by HPLC-ECD, including factors such as different proportions of mobile phase, column temperature and flow rate of the mobile phase. The results showed that 2% acetic acid water mobile phase can obtain the best results, and the gradient elution mode was used to separate the 10 target phenolic acids with methanol and acetic acid water as the mobile phase.

[0076] S14, conducting a determination experiment of the content of polyphenolic compounds, including a preliminary determination experiment and a main determination experiment, wherein the preliminary determination experiment includes a precision experiment, a repeatability experiment and a stability experiment; the main determination experiment is used to calculate a standard curve, and determine the polyphenolic compounds in the sample to be tested based on the standard curve.

[0077] In this embodiment:

[0078] Precision experiment: 10.0ul of the mixed standard solution was accurately weighed and measured 6 times in succession. The RSDs of the retention time and peak area of ​​the eight peaks in the mixed standard solution were less than 0.9% and 1.0%, respectively, and the precision experiment performed well.

[0079] Repeatability experiment: Take 6 portions of sample numbered S1, 10.0ul each, and measure 6 times continuously. The RSD of retention time and peak area are less than 0.81% and 2.3% respectively, and the repeatability is good.

[0080] Stability test: The stability test was carried out at room temperature. Six samples labeled S1 were measured every hour for 12 hours. The RSDs of retention time and peak area were less than 0.76% and 1.5%, respectively, indicating good stability.

[0081] Standard curve: The correlation coefficients of the regression equations of the 10 polyphenol compounds were all greater than 0.999, indicating that there was a strong linear correlation between the content of the ten polyphenol compounds and the chromatographic peak area. The detection limit and quantitative limit that met the chromatographic conditions were obtained when the signal-to-noise ratio S / N was 3 and the signal-to-noise ratio S / N was 10, respectively. The detection limit of each component was 0.0015-0.1202ug / ml, proving that the HPLC-ECD method is very sensitive and can be used to detect many trace components.

[0082] Determination of polyphenolic compound content: Except for rutin, other polyphenolic compounds all have certain contents in sunflower honey. Although Y1, Y2, and Y3 are all rape honey, there are slight differences in their composition and content. This may be due to the difference in vegetation in different regions. Specifically, Y3 does not contain rutin, but Y1 and Y2, which are also rape honey, have higher rutin content. Caffeic acid was not detected in Y1 and Y2, while Y3 has a higher caffeic acid content. Several polyphenolic compounds with higher contents were detected in rape honey (rutin: Y1, kaempferol Y2, Y3), which is the same as what was observed in the chromatogram. Sunflower honey contains more caffeic acid, galangin, and galangin, while rape honey contains more rutin, galangin, and galangin. However, the antioxidant capacity of different polyphenolic compounds is related to their molecular structure, and the total antioxidant activity in honey cannot be determined by the amount of content.

[0083] S2, based on the measured content of polyphenolic compounds, using HPLC-ECD method to establish fingerprints of the same kind of honey from multiple different regions, by analyzing the differences in fingerprints of the same kind of honey, regional calibration of polyphenolic compounds is performed on it to obtain regional calibration values, the regional calibration values ​​are subjected to first similarity and cluster analysis, and a determination model for distinguishing the same kind of honey from different origins is established;

[0084] As a preferred embodiment, the first similarity and cluster analysis to establish a determination model for distinguishing the same type of honey from different origins includes:

[0085] The peak time of the element with the highest content in each sample is in the middle of the entire chromatogram, the peak shape is stable and there is no influence of other peaks, and the peak corresponding to the element is selected as the reference peak, that is, the internal standard peak;

[0086] Calculating the normalized areas of other multiple common peaks based on the reference peak;

[0087] Classifying the samples according to the normalized areas and forming a matrix;

[0088] The similarity between relative peak areas was calculated and classified, and a determination model for distinguishing the same type of honey from different origins was established based on the classification.

[0089] In this embodiment:

[0090] First, the polyphenols in sunflower honey were analyzed, and then the quality of sunflower honey was evaluated by systematically comparing the chromatograms of the samples. The results showed that:

[0091] (1) Quantitative and qualitative analysis of eight antioxidant active substances in sunflower honey using high performance liquid chromatography. This method has the advantages of being rapid, simple, and accurate.

[0092] (2) The contents of the eight phenolic compounds in sunflower honey vary somewhat due to the different sources of the samples;

[0093] (3) Combining similarity and cluster analysis, sunflower honeys from different regions were distinguished. The results showed that genetic and geographical factors may be important factors in determining the characteristic components and chromatographic models of sunflower honey. This method can also be used for quality control and source identification of other crops containing polyphenols during production.

[0094] S3, establishing HPLC-ECD fingerprints of different types of honey, calibrating the types of polyphenol compounds by analyzing the differences in the HPLC-ECD fingerprints of different types of honey to obtain type calibration values, and performing a second similarity and cluster analysis on the type calibration values, thereby establishing a measurement model for distinguishing different types of honey.

[0095] As a preferred embodiment, the step of performing a second similarity and cluster analysis on the type calibration values ​​to establish a measurement model for distinguishing different types of honey includes:

[0096] Analyzing the variation trend of the chromatographic peak areas of samples from different sources and regions to obtain a first analysis result, wherein the first analysis result is used to characterize the variation of the content of polyphenolic compounds in different types of samples;

[0097] Analyzing the sample similarity to obtain a second analysis result, wherein the second analysis result is used to identify the sample type by comparing the sample similarity according to a predetermined condition threshold and a standard spectrum;

[0098] Performing cluster analysis on the obtained type calibration values ​​to obtain a third analysis result, wherein the third analysis result is used to distinguish the origin and type of a large number of samples;

[0099] A determination model for distinguishing different types of honey is established based on the first analysis result, the second analysis result and the third analysis result.

[0100] In this embodiment:

[0101] Through the analysis of the trend of chromatographic peak area changes of honey from three different plant sources and different regions, sample similarity analysis and cluster analysis, it is shown that the use of these three methods can reflect the differences between different types of samples. The trend chart of peak area changes can directly and clearly reflect the changes in the content of different types of honey components, and the types of honey can be distinguished by the trend of peak area changes. Similarity analysis can quickly identify the types of samples by comparing the similarities between samples, but the result of similarity calculation requires a certain condition threshold and an accurate and reliable standard map as a reference. Through cluster analysis, honey can be well distinguished according to origin and type, but this method can only classify large quantities of samples, but cannot qualitatively characterize the samples. The qualitative characterization of samples also needs to be compared with standard maps. The simultaneous use of the three methods provides a basis for the construction of a honey fingerprint database and the establishment of a standard map.

[0102] S4, specify the honey type, establish the fingerprint of adulterated honey after adding different mass fractions of rice syrup into it, analyze the differences in the fingerprints of adulterated honey, determine the polyphenol compounds, and then perform the third similarity and cluster analysis on the measured values ​​to establish an adulterated honey identification model.

[0103] As a preferred embodiment, the method of performing a third similarity and cluster analysis on the measured values ​​to establish an adulterated honey identification model comprises:

[0104] Establish standard fingerprints;

[0105] Collect data from 3 to 32 minutes when the chromatogram is eluted, and calculate the similarity between the sample to be tested and the standard fingerprint. The closer the similarity, the more similar the two samples are, and similar analysis results are obtained;

[0106] The peak time of the element with the highest content in each sample is in the middle of the entire chromatogram, the peak shape is stable and there is no influence of other peaks, and the peak corresponding to the element is selected as the reference peak, that is, the internal standard peak;

[0107] Perform interpolation matching on all common peaks other than the reference peak to be clustered, that is, fill in the missing data with a value of 0, and mark the missing time point with the retention time of the peak position;

[0108] The normalized areas of the common peaks other than the reference peak were calculated, and the obtained results were clustered using SPSS to obtain the cluster analysis results;

[0109] A model for identifying adulterated honey is established based on the similarity analysis results and the cluster analysis results.

[0110] As a preferred embodiment, the establishment of the standard fingerprint spectrum includes:

[0111] The chromatographic information of the sample to be tested is obtained by the HPLC-ECD method;

[0112] The component corresponding to each chromatographic peak is determined according to the retention time recorded in the chromatographic information, and the content of the component can be calculated according to the peak area;

[0113] The common characteristics of the same type of honey, i.e., common peaks, are found through retention time, and the similarity between samples is evaluated through the relative peak areas between the common peaks;

[0114] The map information containing the common characteristics of the species is used as the standard fingerprint map.

[0115] Embodiment 2

[0116] See also Figure 3 This embodiment provides a polyphenol compound determination system based on fingerprint, comprising:

[0117] The polyphenol compound determination module 101 is used to determine the chromatographic conditions for detecting polyphenol compounds in the sample and to determine the content of the polyphenol compounds;

[0118] The first model building module 102 is used to establish fingerprints of the same sample from multiple different regions by using the HPLC-ECD method based on the measured content of polyphenolic compounds, perform regional calibration of polyphenolic compounds on the same sample by analyzing the differences in the fingerprints of the same sample to obtain regional calibration values, perform first similarity and cluster analysis on the regional calibration values, and establish a determination model for distinguishing the same samples from different origins;

[0119] The second model building module 103 is used to establish HPLC-ECD fingerprints of different types of samples, analyze the differences in the HPLC-ECD fingerprints of different types of samples, perform type calibration on the polyphenol compounds, and obtain type calibration values, and perform a second similarity and cluster analysis on the type calibration values, so as to establish a determination model for distinguishing different types of samples;

[0120] The third model building module 104 is used to specify the sample type and build the fingerprint of the adulterated sample after adding different mass fractions of counterfeit products thereto, analyze the differences in the fingerprints of the adulterated samples, measure the polyphenol compounds therein, and then perform the third similarity and cluster analysis on the measured values, so as to build an adulterated sample identification model.

[0121] The present invention also provides a memory storing a plurality of instructions, wherein the instructions are used to implement the method as in the first embodiment.

[0122] like Figure 4 As shown, the present invention also provides an electronic device, including a processor 301 and a memory 302 connected to the processor 301, the memory 302 stores a plurality of instructions, and the instructions can be loaded and executed by the processor so that the processor can execute the method as in the first embodiment.

[0123] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for determining polyphenolic compounds based on fingerprint, characterized in that: include: S1, determining the chromatographic conditions for detecting polyphenolic compounds in a sample, and determining the content of the polyphenolic compounds; S2, based on the determined content of polyphenolic compounds, using HPLC-ECD method to establish fingerprints of the same sample from multiple different regions, by analyzing the differences in the fingerprints of the same sample, regional calibration of polyphenolic compounds is performed on the sample to obtain regional calibration values, the regional calibration values ​​are subjected to first similarity and cluster analysis, and a determination model for distinguishing the same samples from different origins is established; S3, establishing HPLC-ECD fingerprints of different types of samples, performing type calibration of polyphenol compounds on the samples by analyzing the differences in the HPLC-ECD fingerprints of the different types of samples to obtain type calibration values, and performing a second similarity and cluster analysis on the type calibration values, thereby establishing a determination model for distinguishing different types of samples; S4, specify the sample type, establish the fingerprint of the adulterated sample after adding different mass fractions of counterfeit products into it, analyze the differences in the fingerprints of the adulterated samples, measure the polyphenol compounds, and then perform the third similarity and cluster analysis on the measured values ​​to establish the adulterated sample identification model.

2. A method for determining polyphenolic compounds based on fingerprint according to claim 1, characterized in that: The S1 includes: S11, prepare experimental materials and experimental instruments; S12, preparing standard substances and samples to be tested; S13, select and optimize chromatographic conditions; S14, conducting a determination experiment of the content of polyphenol compounds, including a preliminary determination experiment and a main determination experiment.

3. A method for determining polyphenolic compounds based on fingerprint according to claim 2, characterized in that: The preliminary experiments include precision experiments, repeatability experiments and stability experiments; the main experiments are used to calculate the standard curve and determine the polyphenol compounds in the sample to be tested based on the standard curve.

4. A method for determining polyphenolic compounds based on fingerprint according to claim 3, characterized in that: The step of performing a first similarity and cluster analysis on the regional calibration values ​​to establish a determination model for distinguishing the same samples from different origins comprises: The peak time of the element with the highest content in each sample is in the middle of the entire chromatogram, the peak shape is stable and there is no influence of other peaks, and the peak corresponding to the element is selected as the reference peak, that is, the internal standard peak; Calculating the normalized areas of multiple other common peaks based on the reference peak; Classifying the samples according to the normalized areas and forming a matrix; The similarity between relative peak areas was calculated and classified, and a determination model for distinguishing the same samples from different origins was established based on the classification.

5. A method for determining polyphenolic compounds based on fingerprint according to claim 4, characterized in that: The step of performing a second similarity and cluster analysis on the type calibration values ​​to establish a measurement model for distinguishing samples of different types includes: Analyzing the variation trend of the chromatographic peak areas of samples from different sources and regions to obtain a first analysis result, wherein the first analysis result is used to characterize the variation of the content of polyphenolic compounds in different types of samples; Analyzing the sample similarity to obtain a second analysis result, wherein the second analysis result is used to identify the sample type by comparing the sample similarity according to a predetermined condition threshold and a standard spectrum; Performing cluster analysis on the obtained type calibration values ​​to obtain a third analysis result, wherein the third analysis result is used to distinguish the origin and type of a large number of samples; A measurement model for distinguishing different types of samples is established based on the first analysis result, the second analysis result, and the third analysis result.

6. A method for determining polyphenol compounds based on fingerprint according to claim 5, characterized in that: The method of performing a third similarity and cluster analysis on the measured values ​​to establish an adulterated sample identification model comprises: Establish standard fingerprints; Collect data from 3 to 32 minutes when the chromatogram is eluted, and calculate the similarity between the sample to be tested and the standard fingerprint. The closer the similarity, the more similar the two samples are, and similar analysis results are obtained; The peak time of the element with the highest content in each sample is in the middle of the entire chromatogram, the peak shape is stable and there is no influence of other peaks, and the peak corresponding to the element is selected as the reference peak, that is, the internal standard peak; Perform interpolation matching on all common peaks other than the reference peak to be clustered, that is, fill in the missing data with a value of 0, and mark the missing time point with the retention time of the peak position; The normalized areas of the common peaks other than the reference peak were calculated, and the obtained results were clustered using SPSS to obtain the cluster analysis results; A model for identifying adulterated samples is established based on the similarity analysis results and the cluster analysis results.

7. A method for determining polyphenolic compounds based on fingerprint according to claim 6, characterized in that: The establishment of the standard fingerprint spectrum includes: The chromatographic information of the sample to be tested is obtained by the HPLC-ECD method; The component corresponding to each chromatographic peak is determined according to the retention time recorded in the chromatographic information, and the content of the component can be calculated according to the peak area; The commonality of samples of the same type, i.e., common peaks, is found through retention time, and the similarity between samples is evaluated through the relative peak areas between the common peaks; The map information containing the common characteristics of the species is used as the standard fingerprint map.

8. A polyphenol compound determination system based on fingerprint spectrum, used for implementing the method described in any one of claims 1 to 7, characterized in that: include: A polyphenol compound determination module (101) is used to determine the chromatographic conditions for detecting polyphenol compounds in a sample and to determine the content of polyphenol compounds; The first model building module (102) is used to establish fingerprints of the same sample from multiple different regions by using the HPLC-ECD method based on the determined content of polyphenolic compounds, perform regional calibration of polyphenolic compounds on the same sample by analyzing the differences in the fingerprints of the same sample to obtain regional calibration values, perform first similarity and cluster analysis on the regional calibration values, and establish a determination model for distinguishing the same samples from different origins; The second model building module (103) is used to establish HPLC-ECD fingerprints of samples of different types, analyze the differences in the HPLC-ECD fingerprints of samples of different types, perform type calibration on the polyphenol compounds, and obtain type calibration values, and perform a second similarity and cluster analysis on the type calibration values, thereby establishing a determination model for distinguishing samples of different types; The third model building module (104) is used to specify the sample type, establish the fingerprint of the adulterated sample after adding different mass fractions of counterfeit products thereto, analyze the differences in the fingerprint of the adulterated sample, measure the polyphenol compounds thereon, and perform the third similarity and cluster analysis on the measured values, thereby establishing an adulterated sample identification model.

9. An electronic device, characterized in that: The method comprises a processor and a memory, wherein the memory stores a plurality of instructions, and the processor is used to read the instructions and execute the method according to any one of claims 1 to 7.

10. A computer-readable storage medium, characterized in that: The computer-readable storage medium stores a plurality of instructions, and the plurality of instructions can be read by a processor to execute the method according to any one of claims 1 to 7.

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